Compounds for use in the treatment of autoimmune diseases, immuno-allergical diseases and organ or tissue transplantation rejection

ABSTRACT

The present invention provides compounds, pharmaceutical compositions and methods for treating, immuno-allergical diseases, autoimmune diseases, and organ or tissue rejection following transplantation.

RELATED APPLICATIONS

This application is a national stage application, filed under 35 U.S.C.§ 371, of International Application No. PCT/IB2003/004993, filed on Oct.1, 2003, which claims priority from U.S. application Ser. No.60/415,508, filed Oct. 3, 2002.

FIELD OF THE INVENTION

The present invention relates to novel compounds useful in the treatmentof autoimmune diseases, immuno-allergical disease and organ or tissuetransplantation rejection.

BACKGROUND OF THE INVENTION

Diseases of the immune systems pose a major threat due to thepotentially devastating effects that such diseases have on humanity. Oneclass of diseases related to the immune system are immuno-allergicaldiseases. Immuno-allergical diseases are a major cause of concern, andmuch research is being conducted in order to develop potent compoundsthat will be effective in treating such diseases. Allergy is a state ofhypersensitivity induced by exposure to a particular antigen (allergen)resulting in harmful immunologic reaction on subsequent exposures. Anexample of an immuno-allergical disease is allergic rhinitis (hayfever). Allergic rhinitis is a common immuno-allergic condition thataffects one in every five Americans. Over one billion dollars is spenteach year in the U.S. to treat this condition. Sneezing, nasalcongestion, and eye irritation are some of the symptoms of allergicrhinitis. Another example of an immuno-allergical disease is bronchialasthma—a breathing problem that results from spasm (bronchospasm) of themuscles surrounding the walls of the lung airways (bronchi). Allergicasthma is the most common type of asthma, typically first appearing inchildhood.

Another example of an immuno-allergical disease is psoriasis, which is achronic skin disease characterized by scaling and inflammation, and itaffects 1.5 to 2 percent of the United States population, or almost 5million people. It occurs in all age groups and about equally in men andwomen. People with psoriasis may suffer discomfort, restricted motion ofjoints, and emotional distress.

Another type of immuno-allergical disease is Crohn's disease, which is achronic inflammatory disease of the intestines. It primarily causesulceration in the small and large intestines but can affect thedigestive system anywhere between the mouth and the anus. The disease isfound in equal frequency in men and women, and it usually affects youngpatients in their teens or early twenties. Once the disease begins, ittends to be a chronic, recurrent condition with periods of remission anddisease exacerbation.

A third class of diseases related to the immune system are autoimmunediseases. Autoimmune diseases are illnesses that occur when the body'stissues are attacked by its own immune system. The immune system is acomplex organization within the body that is designed normally to seekand destroy invaders of the body, particularly infections. Patients withthese diseases have unusual antibodies in their blood that target theirown body tissues. An example of an autoimmune disease is systemic lupuserythematosus. Lupus is a chronic inflammatory condition, caused byautoimmune disease, which causes disease of the skin, heart, lungs,kidneys, joints, and nervous system. When internal organs are involved,the condition is called systemic lupus erythematosus. Another type of anautoimmune disease is autoimmune thyroiditis, which is an autoimmunedisease of the thyroid. Another type of autoimmune disease is rheumatoidarthritis, which causes chronic inflammation of the joints, the tissuearound the joints, as well as other organs in the body.

Another example of an autoimmune system is experimental autoimmuneenoephalomyelitis (EAE). EAE is an inflammatory condition of the centralnervous system, which is the murine equivalent to multiple sclerosis.

A forth class of conditions related to the immune system is organ ortissue rejection following transplantation. Organ or tissue rejection isa major complication occurring in patients who have undergonetransplantation. For example, chronic graft-versus-host disease (cGvHD),a major complication occurring in patients post-allogeneic bone marrowtransplantation, is believed to be the result of an autoimmune-likeprocess mediated by immunocompetent T-cells. Chronic GvHD often resultsin sclerodermoid-fibrotic skin lesions [see, e.g., Chosidow et al.,Sclerodermatous chronic graft-versus-host disease analysis of sevencases. J Am Acad Dermatol 26:49-53, 1992]. Other examples of organ ortissue rejection following transplantation include skin graft rejectionand cardiac graft rejection.

Autoimmune diseases and immuno-allergical diseases pose a major problemto society. Organ or tissue rejection following transplantation presentsanother problem that severely limits the use and application of tissueand organ transplantation in medicine.

SUMMARY OF THE INVENTION

The present invention relates to novel compounds useful in thetreatment, prevention, and control of immuno-allergical diseases,autoimmune diseases and organ or tissue transplantation rejectionfollowing transplantation.

The present invention provides a compound represented by the structureof formula I

wherein

-   the dotted line represents a single or a double bond;

In one embodiment, R₁ and R₂ are the same or different, andindependently of each other represent —CH₂OH, —CH₂OR₄, —CH(OH)CH₃,—CH(OR₄)CH₃, or a group represented by the formula

where R₄ is a linear or branched C₁-C₄ alkyl; R₅ is H, OH or OR₆ (whereR₆ is a linear or branched C₁-C₄ alkyl); and

-   A-B is a group represented by the formula:

In various embodiments, X is O, —CH₂O, —CH₂CH₂O, —CH(CH₃)CH₂O, or—CH₂CH(CH₃)O, and

-   Z is —CH₂CH₂O, —CH(CH₃)CH₂O, or —CH₂CH(CH₃)O.-   m is an integer of 0 or 1; and n is an integer of 0-50.-   In another embodiment, n is an integer from 1-100.-   In another embodiment, n is an integer from 1-200.-   In yet another embodiment, n is an integer from 1-500.-   In another embodiment, n is an integer from 5-75. For example, n can    be 5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34,    35, 40, 45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12,    17, 34 or 69.

In another embodiment, the present invention provides the salts orhydrates of the compound presented by the structure of formula I.

In one embodiment, R₁ is —CH₂OH, —CH₂OR₄, —CH(OH)CH₃, or —CH(OR₄)CH₃. Inanother embodiment, R₁ is

where R₅ is H or OH. In another embodiment, R₁ is phenyl. In anotherembodiment, R₁ is

In one embodiment, R₂ is —CH₂OH, —CH₂OR₄, —CH(OH)CH₃, or —H(OR₄)CH₃. Inanother embodiment, R₂ is

where R₅ is H or OH. In another embodiment, R₂ is phenyl. In anotherembodiment, R₂ is

In another aspect, the present invention provides a compound representedby the structure of formula II

where

-   the dotted line represents a single or a double bond;-   R₅ and R₅′ are, independently of each other, H, OH or OR₆ where R₆    is a linear or branched C₁-C₄ alkyl.-   In various embodiments, X is O, —CH₂O, —CH₂CH₂O, —CH(CH₃)CH₂O, or    —CH₂CH(CH₃)O; and Z is —CH₂CH₂O, —CH(CH₃)CH₂O, or —CH₂CH(CH₃)O.-   m is an integer of 0 or 1; and n is an integer of 0-50.-   In another embodiment, n is an integer from 1-100.-   In another embodiment, n is an integer from 1-200.-   In yet another embodiment, n is an integer from 1-500.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

In another embodiment, the present invention provides the salts orhydrates of the compound presented by the structure of formula II.

In one embodiment, X is —CH₂O.

In one embodiment, m is 0. In another embodiment, m is 1.

In addition, the present invention provides a compound represented bythe structure of formula III

where the dotted line represents a single or a double bond; R₅ and R₅′are, independently of each other, H, OH or OR₆ (where R₆ is a linear orbranched C₁-C₄ alkyl), Z is —CH₂CH₂O, —CH(CH₃)CH₂O, or —CH₂CH(CH₃)O, andn is an integer of 0-50.

-   In another embodiment, n is an integer from 1-100.-   In another embodiment, n is an integer from 1-200.-   In yet another embodiment, n is an integer from 1-500.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

In another embodiment, the present invention provides the salts orhydrates of the compound presented by the structure of formula III.

In one embodiment, Z is —CH(CH₃)CH₂O.

In addition, the present invention provides a compound represented bythe structure of formula IV

where the dotted line represents a single or a double bond;

-   R₅ and R₅′ are, independently of each other, H, OH or OR₆ (where R₆    is a linear or branched C₁-C₄ alkyl); and-   Z is —CH₂CH₂O, —CH(CH₃)CH₂O, or —CH₂CH(CH₃)O.-   n is an integer of 0-50.-   In another embodiment, n is an integer from 1-100.-   In another embodiment, n is an integer from 1-200.-   In yet another embodiment, n is an integer from 1-500.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

In another embodiment, the present invention provides the salts orhydrates of the compound presented by the structure of formula IV.

In one embodiment, R₅ is H. In another embodiment, R₅ is OH.

In one embodiment, R₅′ is H. In another embodiment, R₅′ is OH.

In one embodiment, n is an integer of 1-20. In another embodiment, n isan integer of 10-20. In another embodiment, n is 17.

In addition, the invention provides a compound of Formula A:

wherein R is a polyalkylene glycol polymer having n units, where n is aninteger from 1-100.

In one embodiment, the polyalkylene glycol polymer is polyisopropyleneglycol.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

In addition, the invention provides a compound of Formula B:

wherein R is a polyalkylene glycol polymer having n units, where n is aninteger from 1-100.

In one embodiment, the polyalkylene glycol polymer is polyisopropyleneglycol.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

In addition, the invention provides a compound of Formula C:

wherein R is a polyalkylene glycol polymer having n units, where n is aninteger from 1-100.

In one embodiment, the polyalkylene glycol polymer is polyisopropyleneglycol.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

In addition, the invention provides a compound of Formula D:

wherein R is a polyalkylene glycol polymer having n units, where n is aninteger from 1-100.

In one embodiment, the polyalkylene glycol polymer is polyisopropyleneglycol.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

In addition, the invention provides a compound of Formula E:

wherein R is a polyalkylene glycol polymer having n units, where n is aninteger from 1-100.

In one embodiment, the polyalkylene glycol polymer is polyisopropyleneglycol.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

In addition, the invention provides a compound of Formula F:

wherein R is a polyalkylene glycol polymer having n units, where n is aninteger from 1-100.

In one embodiment, the polyalkylene glycol polymer is polyisopropyleneglycol.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

Furthermore, in one embodiment, the present invention provides acomposition comprising one or more compounds of formula I, II, III, IV,VII, A, B, C, D, E, or F. In another embodiment, the present inventionprovides a pharmaceutical composition comprising as an active ingredientone or more compounds of formula I, II, III, IV, VII, A, B, C, D, E, orF, together with one or more pharmaceutically acceptable excipients oradjuvants.

Furthermore, in one embodiment, the present invention provides a methodfor the treatment, prevention and control of immuno-allergical diseasesin human as well as veterinary applications, which comprisesadministrating one or more compounds of formula I, II, III, IV, VII, A,B, C, D, E, or F and/or a pharmaceutical composition comprising one ormore compounds of formula I, II, III, IV, VII, A, B, C, D, E, or F. Inone embodiment, the immuno-allergical disease is bronchial asthma,allergic rhinitis, psoriasis or Crohn's disease.

Furthermore, in one embodiment, the present invention provides a methodfor the treatment, prevention and control of autoimmune diseases inhuman as well as veterinary applications, which comprises administratingone or more compounds of formula I, II, III, IV, VII, A, B, C, D, E, orF and/or a pharmaceutical composition comprising one or more compoundsof formula I, II, III, IV, VII, A, B, C, D, E, or F. In one embodiment,the autoimmune disease is systemic lupus erythematosus, autoimmunethyroiditis, rheumatoid arthritis, diabetes, multiple sclerosis andexperimental autoimmune encephalomyelitis.

Furthermore, in one embodiment, the present invention provides a methodfor the treatment, prevention and control of organ or tissuetransplantation rejection in human as well as veterinary applications,which comprises administrating one or more compounds of formula I, II,III, IV, VII, A, B, C, D, E, or F and/or a pharmaceutical compositioncomprising one or more compounds of formula I, II, III, IV, VII, A, B,C, D, E, or F. In one embodiment, the organ or tissue transplantationrejection is kidney rejection, bone marrow rejection, skin graftrejection, cardiac graft rejection or chronic-graft-versus-host disease.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described below. All publications, patent applications,patents, and other references mentioned herein are incorporated byreference in their entirety. In the case of conflict, the presentspecification, including definitions, will control. In addition, thematerials, methods, and examples are illustrative only and are notintended to be limiting.

The above description sets forth rather broadly the more importantfeatures of the present invention in order that the detailed descriptionthereof that follows may be understood, and in order that the presentcontributions to the art may be better appreciated. Other objects andfeatures of the present invention will become apparent from thefollowing detailed description considered in conjunction with theaccompanying drawings. It is to be understood, however, that thedrawings are designed solely for the purposes of illustration and not asa definition of the limits of the invention, for which reference shouldbe made to the appended claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a bar graph depicting the effect of AV 75 on spleencells±ConA.

FIG. 2 is a bar graph depicting the effect of AV 77 on spleencells±ConA.

FIG. 3 is a bar graph depicting the effect of AV 75±AV 77 on spleencells±ConA.

FIG. 4 is a bar graph summarizing the effect of AV 75 and AV 77 onspleen cells±ConA.

FIG. 5 is a bar graph depicting the effect of AV 75 on spleencells±ConA.

FIG. 6 is a bar graph depicting the effect of AV 75 on spleencells±ConA.

FIG. 7 is a bar graph depicting the effect of AV 81 on spleencells±ConA.

FIG. 8 is a bar graph depicting the effect of AV 82 on spleencells±ConA.

FIG. 9 is a bar graph depicting the effect of AV 86 on spleencells±ConA.

FIG. 10 is a bar graph depicting the effect of AV 87 on spleencells±ConA.

FIG. 11 is a bar graph depicting the effect of AV 75 on spleencells±PHA.

FIG. 12 is a bar graph depicting the effect of AV 75 on spleencells±PHA.

FIG. 13 is a bar graph depicting the effect of AV 76 on spleencells±PHA.

FIG. 14 is a bar graph depicting the effect of AV 77 on spleencells±PHA.

FIG. 15 is a bar graph depicting the effect of AV 74 on PMBCs±PHA.

FIG. 16 is a bar graph depicting the effect of AV 75 on PMBCs±PHA

FIG. 17 is a bar graph depicting the effect of AV 76 on PMBCs±PHA.

FIG. 18 is a bar graph depicting the effect of AV 77 on PMBCs±PHA.

FIG. 19 is a bar graph depicting the effect of AV 72 and AV 73 onPMBCs±ConA relative to those treated with cyclosporin A.

FIG. 20 is a bar graph depicting the effect of AV 74 on PMBCs±ConArelative to those treated with cyclosporin A.

FIG. 21 is a bar graph depicting the effect of AV 75 on PMBCs±ConArelative to those treated with cyclosporin A.

FIG. 22 is a bar graph depicting the effect of AV 76 on PMBCs±ConArelative to those treated with cyclosporin A.

FIG. 23 is a bar graph depicting the effect of AV 77 on PMBCs±ConArelative to those treated with cyclosporin A.

FIG. 24 is a bar graph depicting the effect of AV 61, AV75, and AV 77 onPMBCs±ConA relative, to those treated with cyclosporin A.

FIG. 25 is a bar graph depicting the effect of AV 61, AV75, and AV 77 ondendritic cells±ConA relative to those treated with cyclosporin A.

FIGS. 26A, B, and C are line graphs depicting the effect of AV 75 onexperimental autoimmune encephalomyelitis. The pathological score isrecorded as a function of days after treatment.

FIG. 27A is a bar graph summarizing the effect of AV 75 on theproliferative response to a variety of antigens by lymphocytes obtainedfrom mouse lymph nodes; FIG. 27B depicts the effect of AV 75 on theproliferative response to LPS by lymphocytes obtained from mouse lymphnodes; FIG. 27C depicts the effect of AV 75 on the proliferativeresponse to SEB by lymphocytes obtained from mouse lymph nodes; FIG. 27Ddepicts the effect of AV 75 on the proliferative response to ConA bylymphocytes obtained from mouse lymph nodes; FIG. 27E depicts the effectof AV 75 on the proliferative response to PPD by lymphocytes obtainedfrom mouse lymph nodes; FIG. 27F depicts the effect of AV 75 on theproliferative response to PLP by lymphocytes obtained from mouse lymphnodes; FIG. 27G depicts the effect of AV 75 on the proliferativeresponse to control treated lymphocytes obtained from mouse lymph nodes.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel compounds and pharmaceuticalcompositions useful in the treatment of immuno-allergical diseases,autoimmune diseases and organ or tissue transplantation rejection. Thepresent invention further provides a method for the treatment,prevention and control of immuno-allergical diseases, autoimmunediseases, and organ or issue transplantation rejection comprisingadministering to a subject one or more of the compounds represented bythe structure of formula I, II, III, IV, VII, A, B, C, D, E, or F.

In one embodiment, the present invention provides a compound representedby the structure of formula I:

wherein

-   R₁ and R₂ are the same or different, and independently of each other    represent —CH₂OH, —CH₂OR₄, —CH(OH)CH₃, —CH(OR₄)CH₃, or a group    represented by the formula

wherein

-   R₄ is a linear or branched C₁-C₄ alkyl;-   R₅ is H, OH or OR₆ wherein R₆ is a linear or branched C₁-C₄ alkyl;-   A-B is a group represented by the formula:

-   X is O, —CH₂O, —CH₂CH₂O, —CH(CH₃)CH₂O, or —CH₂CH(CH₃)O;-   Z is —CH₂CH₂O, —CH(CH₃)CH₂O, or —CH₂CH(CH₃)O;-   m is an integer of 0 or 1; and-   n is an integer of 0-50;-   and salts or hydrates thereof.-   In another embodiment, n is an integer from 1-100.-   In another embodiment, n is an integer from 1-200.-   In yet another embodiment, n is an integer from 1-500.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

In another embodiment, the present invention further provides a compoundrepresented by the structure of formula II:

wherein

-   R₅ and R₅′ are independently of each other H, OH or OR₆ wherein R₆    is a linear or branched C₁-C₄ alkyl;-   X is O, —CH₂O, —CH₂CH₂O, —CH(CH₃)CH₂O, or —CH₂CH(CH₃)O;-   Z is —CH₂CH₂O, —CH(CH₃)CH₂O, or —CH₂CH(CH₃)O;-   m is an integer of 0 or 1; and-   n is an integer of 0-50;-   and salts or hydrates thereof.-   In another embodiment, n is an integer from 1-100.-   In another embodiment, n is an integer from 1-200.-   In yet another embodiment, n is an integer from 1-500.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

In another embodiment, the present invention provides a compoundrepresented by the structure of formula III:

wherein

-   R₅ and R₅′ are independently of each other H, OH or OR₆ wherein R₆    is a linear or branched C₁-C₄ alkyl;-   Z is —CH₂CH₂O, —CH(CH₃)CH₂O, or —CH₂CH(CH₃)O;-   n is an integer of 0-50;-   and salts or hydrates thereof.-   In another embodiment, n is an integer from 1-100.-   In another embodiment, n is an integer from 1-200.-   In yet another embodiment, n is an integer from 1-500.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

The present invention provides a compound represented by the structureof formula IV:

Wherein

-   R₅ and R₅′ are independently of each other H, OH or OR₆ wherein R₆    is a linear or branched C₁-C₄ alkyl;-   n is an integer of 0-50;-   and salts or hydrates thereof.-   In another embodiment, n is an integer from 1-100.-   In another embodiment, n is an integer from 1-200.-   In yet another embodiment, n is an integer from 1-500.

In another embodiment, n is an integer from 5-75. For example, n can be5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40,45, 50, 60, 65, 68, 69, 70, or 75. Preferably, n is, 7, 12, 17, 34 or69.

As contemplated herein, an “alkyl” group refers to a saturated aliphatichydrocarbon, including straight-chain, branched-chain and cyclic alkylgroups. In one embodiment, the alkyl group has 1-4 carbons. In anotherembodiment, the alkyl group is a methyl group. In another embodiment,the alkyl group is an ethyl group. In another embodiment, the alkylgroup is a propyl group. In another embodiment, the alkyl group is abutyl group. The alkyl group may be unsubstituted or substituted by oneor more groups selected from halogen, hydroxy, alkoxy carbonyl, amido,alkylamido, dialkylamido, nitro, amino, alkylamino, dialkylamino,carboxyl, thio and thioalkyl.

In one embodiment, R₁ and R₂ are the same or different in the compoundsof the invention, and, independently of each other, represent —CH₂OH,—CH₂OR₄, —CH(OH)CH₃, —CH(OR₄)CH₃, or a group represented by the formula:

In one embodiment, R₄ is a linear or branched C₁-C₄ alkyl.

In one embodiment, A-B is a group represented by the formula:

In one embodiment, X is O, —CH₂O, —CH₂CH₂O, —CH(CH₃)CH₂O, or—CH₂CH(CH₃)O.

In one embodiment, m is an integer of 0 or 1.

In one embodiment, Z is —CH₂CH₂O, —CH(CH₃)CH₂O, or —CH₂CH(CH₃)O.

In one embodiment, n is an integer of 0-50. In another embodiment, n isan integer of 1-20. In another embodiment, n is an integer of 10-20. Inanother embodiment, n is 17. In various other embodiments, n is 7, 12,or 34. In other embodiments, n is an integer from 1-100, an integer from1-200, an integer from 1-500. In another embodiment, n is an integerfrom 5-75. For example, n can be 5, 6, 7, 8, 10, 11, 12, 13, 15, 16, 17,18, 20, 25, 30, 33, 34, 35, 40, 45, 50, 60, 65, 68, 69, 70, or 75.Preferably, n is, 7, 12, 17, 34 or 69.

In one embodiment, R₅ is H, OH or OR₆.

In one embodiment, R₅′ is H, OH or OR₆.

In one embodiment, R₆ is a linear or branched C₁-C₄ alkyl.

The synthetic methodologies for obtaining the compounds are disclosed indetail in the Examples section, below. However, it should be apparent toa person skilled in the art that the compounds of the present inventioncan be prepared by any feasible synthetic method and that the synthesesset forth in the Experimental Details Section are in no way limiting.Various synthetic methods for the preparation of these compounds will beknown to a person skilled in the art. Compounds of the invention may befurther modified as allowed by the rules of chemistry. Suchmodifications include the addition of various substituents (e.g.,hydroxylation, carboxylation, methylation, etc.), generation ofenantiomers, creation of acid- or base-addition salts, and the like.Other modifications include adding polyalkylene glycol polymers.

The compounds of the invention may be synthesized as polyalkylene glycol(PAG) conjugates. Typical polymers used for conjugation includepoly(ethylene glycol) (PEG), also known as or poly(ethylene oxide) (PEO)and polypropylene glycol (including poly isopropylene glycol). Theseconjugates are often used to enhance solubility and stability and toprolong the blood circulation half-life of molecules.

In its most common form, a polyalkylene glycol (PAG), such as PEG is alinear polymer terminated at each end with hydroxyl groups:HO—CH₂CH₂O—(CH₂CH₂O)_(n)—CH₂CH₂—OH.

The above polymer, alpha-, omega-dihydroxylpoly(ethylene glycol), canalso be represented as HO-PEG-OH, where it is understood that the-PEG-symbol represents the following structural unit:—CH₂CH₂O—(CH₂CH₂O)_(n)—CH₂CH₂—where n typically ranges from about 4 to about 10,000. PEG is commonlyused as methoxy-PEG-OH, or mPEG, in which one terminus is the relativelyinert methoxy group, while the other terminus is a hydroxyl group thatis subject to ready chemical modification. Additionally, random or blockcopolymers of different alkylene oxides (e.g., ethylene oxide andpropylene oxide) that are closely related to PEG in their chemistry canbe substituted for PEG in many of its applications.

PAGs are polymers which typically have the properties of solubility inwater and in many organic solvents, lack of toxicity, and lack ofimmunogenicity. One use of PAGs is to covalently attach the polymer toinsoluble molecules to make the resulting PAG-molecule “conjugate”soluble. For example, it has been shown that the water-insoluble drugpaclitaxel, when coupled to PEG, becomes water-soluble. Greenwald, etal., J. Org. Chem., 60:331-336 (1995).

Polyalkylated compounds of the invention typically contain between 1 and500 monomeric units. Other PAG compounds of the invention containbetween 1 and 200 monomeric units. Still other PAG compounds of theinvention contain between 1 and 100 monomeric units. For example, thepolymer may contain 1, 10, 20, 30, 40, 50, 55, 60, 65, 70, 75, 80, 85,90, 95, or 100 monomeric units. Some compounds of the invention containpolymers which include between 5 and 75 or between 1 and 50 monomericunits. For example, the polymer may contain 2, 3, 5, 6, 7, 8, 10, 11,12, 13, 15, 16, 17, 18, 20, 25, 30, 33, 34, 35, 40, 45, 50, 60, 65, 68,69, 70, or 75 monomeric units. Preferably, n is, 7, 12, 17, 34 or 69.The polymers can be linear or branched.

It is to be understood that compounds which have been modified by theaddition of a PAG moiety may include a mixture of polymers which have avarying number of monomeric units. Typically, the synthesis of aPAG-modified compound (e.g., a PAG-conjugate) will produce a populationof molecules with a Poisson distribution of the number of monomericunits per polymer in the conjugate. Thus, a compound described as havinga polymer of N=7 monomeric units refers not only to the actual polymersin that population being described as having N=7 monomeric units, butalso to a population of molecules with the peak of the distributionbeing 7. The distribution of monomeric units in a given population canbe determined, e.g., by nuclear magnetic resonance (NMR) or by massspectrometry (MS).

Throughout this application, conventional terminology is used todesignate the isomers as described below and in appropriate text booksknown to those of ordinary skill in the art. (see, e.g., Principles inBiochemistry, Lehninger (ed.), page 99-100, Worth Publishers, Inc.(1982) New York, N.Y.; Organic Chemistry, Morrison and Boyd, 3rdEdition, Chap. 4, Allyn and Bacon, Inc., Boston, Mass. (1978).

Certain compounds of the present invention may exist in particulargeometric or stereoisomeric forms. The present invention contemplatesall such compounds, including cis- and trans-isomers, R- andS-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemicmixtures thereof, and other mixtures thereof, as falling within thescope of the invention. Additional asymmetric carbon atoms may bepresent in a substituent such as an alkyl group. All such isomers, aswell as mixtures thereof, are intended to be included in this invention.

A carbon atom which contains four different substituents is referred toas a chiral center. A chiral center can occur in two different isomericforms. These forms are identical in all chemical and physical propertieswith one exception, the direction in which they can cause the rotationof plane-polarized light. These compounds are referred to as being“optically active,” i.e., the compounds can rotate the plane-polarizedlight in one direction or the other.

The four different substituent groups attached to a carbon can occupytwo different arrangements in space. These arrangements are notsuperimposable mirror images of each other and are referred to asoptical isomers, enantiomers, or stereoisomers. A solution of onestereoisomer of a given compound will rotate plane polarized light tothe left and is called the levorotatory isomer [designated (−)]; theother stereoisomer for the compound will rotate plane polarized light tothe same extent but to the right and is called dextrorotatory isomer[designated (+)].

The R S system was invented to avoid ambiguities when a compoundcontains two or more chiral centers. In general, the system is designedto rank the four different substituent atoms around an asymmetric carbonatom in order of decreasing atomic number or in order of decreasingvalance density when the smallest or lowest-rank group is pointingdirectly away from the viewer. The different rankings are well known inthe art and are described on page 99 of Lehninger. If the decreasingrank order is seen to be clock-wise, the configuration around the chiralcenter is referred to as R; if the decreasing rank order iscounter-clockwise, the configuration is referred to as S. Each chiralcenter is named accordingly using this system.

If, for instance, a particular enantiomer of a compound of the presentinvention is desired, it may be prepared by asymmetric synthesis, or byderivation with a chiral auxiliary, where the resulting diastereomericmixture is separated and the auxiliary group cleaved to provide the puredesired enantiomers. Alternatively, where the molecule contains a basicfunctional group, such as amino, or an acidic functional group, such ascarboxyl, diastereomeric salts are formed with an appropriateoptically-active acid or base, followed by resolution of thediastereomers thus formed by fractional crystallization orchromatographic means well known in the art, and subsequent recovery ofthe pure enantiomers.

The compositions and pharmaceutical compositions of the presentinvention comprise one or more of the compounds of the presentinvention, either in a pure form or a partially pure form. Similarly,the methods of the present invention comprise using one or morecompounds, wherein the compounds are in a pure form, a partially pureform.

In one embodiment, a composition of the invention comprises at least oneof the compounds of the present invention, i.e. one or more of thecompounds represented by the structures of formula I, II, III, IV, VII,A, B, C, D, E, or F. In another embodiment, a composition of theinvention comprises a mixture of at least two of the compoundsrepresented by the structures of formula I, II, III, IV, VII, A, B, C,D, E, or F. In another embodiment, a composition of the inventioncomprises a mixture of at least five of the compounds represented by thestructures of formula I, II, III, IV, VII, A, B, C, D, E, or F. Inanother embodiment, a composition of the invention comprises a mixtureof at least ten of the compounds represented by the structures offormula I, II, III, IV, VII, A, B, C, D, E, or F.

It has now been surprisingly found that one or more compoundsrepresented by the structures of formula I, II, III, IV, VII, A, B, C,D, E, or F are effective against a very wide variety ofimmuno-allergical diseases and conditions. Immuno-allergical diseasesare allergy-associated diseases of the immune system. Nonlimitingexamples of immuno-allergical diseases against which the compounds ofthe present invention are active are bronchial asthma, allergicrhinitis, psoriasis and Crohn's disease.

Thus, in one embodiment, the present invention provides a method for thetreatment, prevention and control of immuno-allergical diseases in humanas well as veterinary applications. In one embodiment, the methodcomprises administering to a subject one or more compounds representedby the structures of formula I, II, III, IV, VII, A, B, C, D, E, or F.In another embodiment, the method comprises administering to a subject apharmaceutical composition comprising one or more compounds representedby the structures of formula I, II, III, IV, VII, A, B, C, D, E, or F.

It has now further been surprisingly found that one or more compoundsrepresented by the structures of formula I, II, III, IV, VII, A, B, C,D, E, or F are effective against a very wide variety of autoimmunediseases and conditions. Autoimmune diseases are illnesses that occurwhen the body tissues are mistakenly attacked by its own immune system.The immune system is a complex organization of cells and antibodiesdesigned normally to “seek and destroy” invaders of the body,particularly infections. Patients with these diseases have antibodies intheir blood that target their own body tissues, where they can beassociated with inflammation. Nonlimiting examples of autoimmunediseases against which the compounds of the present invention are activeare systemic lupus erythematosus, autoimmune thyroiditis, rheumatoidarthritis, diabetes, multiple sclerosis and experimental autoimmuneencephalomyelitis.

Thus, in one embodiment, the present invention provides a method for thetreatment, prevention and control of autoimmune diseases in human aswell as veterinary applications. In one embodiment, the method comprisesadministering to a subject one or more compounds represented by thestructures of formula I, II, III, IV, VII, A, B, C, D, E, or F. Inanother embodiment, the method comprises administering to a subject apharmaceutical composition comprising one or more compounds representedby the structures of formula I, II, III, IV, VII, A, B, C, D, E, or F.

It has now further been surprisingly found that one or more compoundsrepresented by the structures of formula I, II, III, IV, VII, A, B, C,D, E, or Fare effective against organ or tissue transplantationrejection. In one embodiment, the organ or tissue transplantationrejection is bone-marrow rejection, skin graft rejection, cardiac graftrejection or chronic-graft-versus-host disease.

Thus in one embodiment, the present invention provides a method for thetreatment, prevention and control of organ or tissue transplantationrejection in human as well as veterinary applications. In oneembodiment, the method comprises administering to a subject one or morecompounds represented by the structures of formula I, II, III, IV, VII,A, B, C, D, E, or F. In another embodiment, the method comprisesadministering to a subject a pharmaceutical composition comprising oneor more compounds represented by the structures of formula I, II, III,IV, VII, A, B, C, D, E, or F.

Methods of administration are well known to a person skilled in the art.Methods of administration include but are not limited to parenterally,transdermally, intramuscularly, intravenously, intradermally,intranasally, subcutaneously, intraperitonealy, or intraventricularly orrectally. Methods and means of administration are known to those skilledin the art. For example, U.S. Pat. Nos. 5,693,622; 5,589,466; 5,580,859;and 5,566,064, which are hereby incorporated by reference in theirentirety.

In addition, the present invention provides a pharmaceutical compositioncomprising as an active ingredient one or more compounds of the presentinvention, together with one or more pharmaceutically acceptableexcipients. As used herein, “pharmaceutical composition” can mean atherapeutically effective amount of one or more compounds of the presentinvention together with suitable excipients and/or carriers useful forthe treatment of immuno-allergical diseases, autoimmune diseases, andorgan or tissue transplantation rejection. A “therapeutically effectiveamount” as used herein refers to that amount that provides a therapeuticeffect for a given condition and administration regimen. Suchcompositions can be administered by any one of the methods listedhereinabove.

A further aspect of the invention comprises a compound of the inventionin combination with other compounds of the invention. A compound of theinvention may also be administered in combination with ananti-inflammatory agent, an immunosuppressant, an antiviral agent, orthe like. Furthermore, the compounds of the invention may beadministered in combination with a chemotherapeutic agent such as analkylating agent, anti-metabolite, mitotic inhibitor or cytotoxicantibiotic, as described above. In general, the currently availabledosage forms of the known therapeutic agents for use in suchcombinations will be suitable.

“Combination therapy” (or “co-therapy”) includes the administration of acompound of the invention and at least a second agent as part of aspecific treatment regimen intended to provide the beneficial effectfrom the co-action of these therapeutic agents. The beneficial effect ofthe combination includes, but is not limited to, pharmacokinetic orpharmacodynamic co-action resulting from the combination of therapeuticagents. Administration of these therapeutic agents in combinationtypically is carried out over a defined time period (usually minutes,hours, days or weeks depending upon the combination selected).“Combination therapy” may, but generally is not, intended to encompassthe administration of two or more of these therapeutic agents as part ofseparate monotherapy regimens that incidentally and arbitrarily resultin the combinations of the present invention.

“Combination therapy” is intended to embrace administration of thesetherapeutic agents in a sequential manner, that is, wherein eachtherapeutic agent is administered at a different time, as well asadministration of these therapeutic agents, or at least two of thetherapeutic agents, in a substantially simultaneous manner.Substantially simultaneous administration can be accomplished, forexample, by administering to the subject a single capsule having a fixedratio of each therapeutic agent or In multiple, single capsules for eachof the therapeutic agents.

Sequential or substantially simultaneous administration of eachtherapeutic agent can be effected by any appropriate route including,but not limited to, oral routes, intravenous routes, intramuscularroutes, and direct absorption through mucous membrane tissues. Thetherapeutic agents can be administered by the same route or by differentroutes. For example, a first therapeutic agent of the combinationselected may be administered by intravenous injection while the othertherapeutic agents of the combination may be administered orally.Alternatively, for example, all therapeutic agents may be administeredorally or all therapeutic agents may be administered by intravenousinjection. The sequence in which the therapeutic agents are administeredis not narrowly critical.

“Combination therapy” also can embrace the administration of thetherapeutic agents as described above in further combination with otherbiologically active ingredients and non-drug therapies (e.g., surgery orradiation treatment.) Where the combination therapy further comprises anon-drug treatment, the non-drug treatment may be conducted at anysuitable time so long as a beneficial effect from the co-action of thecombination of the therapeutic agents and non-drug treatment isachieved. For example, in appropriate cases, the beneficial effect isstill achieved when the non-drug treatment is temporally removed fromthe administration of the therapeutic agents, perhaps by days or evenweeks.

The compounds of the invention and the other pharmacologically activeagent may be administered to a patient simultaneously, sequentially orin combination. It will be appreciated that when using a combination ofthe invention, the compound of the invention and the otherpharmacologically active agent may be in the same pharmaceuticallyacceptable carrier and therefore administered simultaneously. They maybe in separate pharmaceutical carriers such as conventional oral dosageforms which are taken simultaneously. The term “combination” furtherrefers to the case where the compounds are provided in separate dosageforms and are administered sequentially.

The compositions and combination therapies of the invention may beadministered in combination with a variety of pharmaceutical excipients,including stabilizing agents, carriers and/or encapsulation formulationsas described herein.

In one embodiment, the compositions of the present invention areformulated as oral or parenteral dosage forms, such as uncoated tablets,coated tablets, pills, capsules, powders, granulates, dispersions orsuspensions. In another embodiment, the compositions of the presentinvention are formulated for intravenous administration. In anotherembodiment, the compounds of the present invention are formulated inointment, cream or gel form for transdermal administration. In anotherembodiment, the compounds of the present invention are formulated as anaerosol or spray for nasal application. In another embodiment, thecompositions of the present invention are formulated in a liquid dosageform. Examples of suitable liquid dosage forms include solutions orsuspensions in water, pharmaceutically acceptable fats and oils,alcohols or other organic solvents, including esters, emulsions, syrupsor elixirs, solutions and/or suspensions.

Suitable excipients and carriers can be solid or liquid and the type isgenerally chosen based on the type of administration being used.Liposomes may also be used to deliver the composition. Examples ofsuitable solid carriers include lactose, sucrose, gelatin and agar. Oraldosage forms may contain suitable binders, lubricants, diluents,disintegrating agents, coloring agents, flavoring agents, flow-inducingagents, and melting agents. Liquid dosage forms may contain, forexample, suitable solvents, preservatives, emulsifying agents,suspending agents, diluents, sweeteners, thickeners, and melting agentsParenteral and intravenous forms should also include minerals and othermaterials to make them compatible with the type of injection or deliverysystem chosen.

This invention is further illustrated in the Examples section, whichfollows. This section is set forth to aid in an understanding of theinvention but is not intended to, and should not be construed to, limitin any way the invention as set forth in the claims that followthereafter.

EXAMPLES Example 1 Synthesis of Compounds

Compounds of the invention were synthesized and characterized asdescribed below.

0.66 gr, 4 mM, 4-hydroxy hydrocinnamic acid and 4 ml thionyl chloride in30 ml cyclohexane were refluxed for 2 hours. Evaporation gave a yellowsolid to which were added 0.9 gr, 4 mM, phenyl alanine ethyl ester HCl,30 ml dichloromethane and 1 ml triethyl amine. After stirring 2 hours atroom temperature, water and KOH were added to neutral pH and thereaction extracted with dichloromethane Evaporation gave a light yellowviscous oil, which was triturated and recrystallyzed with ethanol togive 0.25 gr, 18%, white solid, mp-213.

NMR CDCl₃ 7.30-6.9 (9H, m), 4.20 (2H, q, J=7.0 Hz), 3.30 (1H, m) 3.10(2H, t, J=7.2 Hz) 2.90 (2H, m), 2.60 (2H, t, J=7.2 Hz), 1.35 (3H, t,J=7.0 Hz). MS-341 M⁺, 10%), 277(15), 194(20), 165(M-phenethyl ester,100%), 149(65) m/e.

0.66 gr, 4 mM, 4-hydroxy hydrocinnamic acid and 4 ml thionyl chloride in30 ml cyclohexane were refluxed for 2 hours. Evaporation gave lightyellow solid to which were added 0.5 gr, 4.1 mM, phenethyl amine, 30 mldichloromethane and 0.6 ml triethyl amine. After stirring for 2 hours atroom temperature, water and KOH were added to neutral pH and thereaction was extracted with dichloromethane. Evaporation gave a viscousoil which was recrystallyzed with ethanol to give 0.3 gr white solid,28%, mp-165.

NMR acetone d₆ 7.35-6.75 (9H, m), 3.40 (2H, q, J=7.1 Hz), 2.90 (2H, t,J=7.2 Hz) 2.75 (2H, t, J=7.2 Hz), 2.42 (2H, t, J=7.1 Hz). Phenethylamine-NMR acetone d₆ 7.2 (5H, m), 2.96 (2H, t, J=7.2 Hz) 2.75 (2H, t,J=7.2 Hz). MS-269(M⁺, 100%), 178(M-benzyl) m/e.

0.66 gr, 4 mM, 4-hydroxy hydrocinnamic acid and 4 ml thionyl chloride in30 ml cyclohexane were refluxed for 1.5 hours. Evaporation gave a lightyellow solid to which were added 0.5 gr, 4.1 mM, histidine amine, 30 mldichloromethane and 0.5 ml triethyl amine. After stirring 2 hours atroom temperature, water and KOH were added to neutral pH and thereaction was extracted with dichloromethane. Evaporation gave a viscousoil which was recrystallyzed with ethanol to give 0.15 gr white solid,15%, mp-245.

NMR acetone d₆ 7.35-(6H, m), 3.42 (2H, q, J=7.1 Hz), 2.93 (2H, t, J=7.2Hz), 2.73 (2H, t, J=7.2 Hz), 2.45 (2H, t, J=7.1 Hz). MS-259(M⁺, 17%),239(25), 213(18), 194(100%), 185(37) m/e

3.2 gr DL phenyl alanine, 20 ml ethylene glycol and 7 ml thionylchloride were refluxed for 2 hours. Workup as above gave 1.3 gr oilwhich was used in the synthesis of AV 28.

NMR acetone d₆ 7.35-(5H, m), 4.50, 3.27, 2.90 (3H, 12 line ABX), 4.32(2H, t, J=7.0 Hz), 3.76 (2H, t, J=7.0 Hz).

1 gr, 6 mM, 4-hydroxy hydrocinnamic acid and 5 ml thionyl chloride in 30ml cyclohexane were refluxed for 1.5 hours. Evaporation gave a lightyellow solid to which were added 1.2 gr AV 27 in 30 ml dichloromethaneand 1 ml triethyl amine. After stirring 2 hours at room temperature,water and KOH were added to neutral pH and the reaction was extractedwith dichloromethane. Evaporation gave a viscous oil which wasrecrystallyzed with ethanol to give 0.18 gr white solid, 8%, mp-224.

NMR acetone d₆ 7.35-6.8 (9H, m), 3.73-2.50 (12H, m).

0.22 gr, 1.3 mM, 4-hydroxy hydrocinnamic acid and 2 ml thionyl chloridein 30 ml cyclohexane were refluxed for 1.5 hours. Evaporation gave alight yellow solid to which were added 0.2 gr, 1.4 mM, tryptamine in 30ml dichloromethane and 0.3 ml triethyl amine. After stirring 1.5 hoursat room temperature, water and KOH were added to neutral pH and thereaction was extracted with dichloromethane. Evaporation gave a viscousoil which was recrystallyzed with ethanol to give 0.11 gr white solid,27%, mp-136.

NMR acetone d₆ 7.36 (2H, d, J=7.8 Hz), 7.0 (8H, m), 3.48 (2H, q, J=7.1Hz), 3.05 (2H, t, J=7.1 Hz), 2.88 (2H, t, J=7.1 Hz), 2.52 (2H, t, J=7.1Hz).

0.22 gr, 1.3 mM, 4-hydroxy hydrocinnamic acid and 2 ml thionyl chloridein 30 ml cyclohexane were refluxed for 1.5 hours. Evaporation gave lightyellow solid to which were added 0.2 gr, 1.5 mM, tyramine, 30 mldichloromethane and 0.3 ml triethyl amine. After stirring for 2 hours atroom temperature, water and KOH were added to neutral pH and thereaction was extracted with dichloromethane. Evaporation gave a viscousoil which was recrystallyzed with ethanol to give 85 mg white solid,23%.

NMR acetone d₆ 7.36 (4H, ABq, J=8.8 Hz), 7.20 (4H, Abq, J=8.6 Hz), 3.48(2H, q, J=7.1 Hz), 3.05 (2H, t, J=7.1 Hz), 2.88 (2H, t, J=7.1 Hz), 2.52(2H, t, J=7.1 Hz).

A. 0.8 gr 4-hydroxy cinnamic acid in 40 ml methanol and 10 drops HClwere refluxed for 12 hours. Workup as above gave 0.6 gr oil, 68% yield.

NMR CDCl₃ 7.02, 6.75 (4H, Abq, J=8.6 Hz), 3.66 (3H, s), 2.86 (2H, t,J=7.4 Hz), 2.60 (2H, t, J=7.4 Hz).

B. 0.6 gr, 3.3 mM, ester from step A and 0.26 gr, 4.2 mM, ethanol aminewere heated at 100 for 3 hours in an open vessel. Chromatography gave0.3 gr recovered ester followed by amide. The viscous oil was trituratedwith acetone-methylene chloride and filtered to give 160 mg white solid,23% yield, mp-102.

NMR acetone d₆ 8.10 (1H, s, OH), 7.03, 6.74 (4H, Abq, J=8.8 Hz), 3.90(1H, t, J=5.2 Hz, NH), 3.54 (2H, q, J=7.1 Hz), 3.28 (2H, t, J=7.1 Hz),2.80 (2H, t, J=8.2 Hz), 2.41 (2H, t, J=8.2 Hz).

0.9 gr, 6 mM, hydrocinnamic acid and 0.6 gr, 6 equivalents, triphosgenin 30 ml dichloromethane and 1.5 ml triethyl amine were stirred 10minutes at room temperature and 0.7 gr phenethyl amine were added. After2 hours at room temperature, workup (HCl) gave a viscous oil which wasrecrystalyzed with hexane-methylene chloride to give 166 mg white solid,11%, mp-91.

NMR acetone d₆ 7.35 (10H, m), 3.40 (2H, q, J=7.2 Hz), 2.90 (2H, t, J=7.4Hz), 2.74 (2H, t, J=7.2 Hz), 2.46 (2H, t, J=7.4 Hz).

Prepared as AV 33, in the same amount but with tyramine instead ofphenethyl amine. Chromatography and trituration with benzene-hexane gave220 mg white solid, 14%, mp-98.

NMR acetone d₆ 7.25 (5H, m), 6.96, 6.75 (4H, Abq, J=8.4 Hz), 3.43 (2H,q, J=6.8 Hz), 2.94 (2H, t, J=7.6 Hz), 2.65 (2H, t, J=6.8 Hz), 2.42 (2H,t, J=7.6 Hz).

Prepared as AV 33, 3 mM, from indole propionic acid and tryptamine.Chromatography and trituration with ethanol gave 162 mg white solid,16%, mp-142.

NMR acetone d₆ 7.57 (2H, d, J=7.8 Hz), 7.36 (2H, d, J=7.8 Hz), 7.0 (8H,m), 3.48 (2H, q, J=7.1 Hz), 3.05 (2H, t, J=7.1 Hz), 2.88 (2H, t, J=7.1Hz), 2.52 (2H, t, J=7.1 Hz).

Prepared as AV 33, 2 mM, from indole propionic acid and phenethyl amine.Chromatography and trituration with ethanol gave 220 mg white viscousoily solid, 37%.

NMR acetone d₆ 7.57 (2H, d, J=7.8 Hz)), 7.25-6.97 (9H, m), 3.44 (2H, q,J=7.1 Hz), 3.10 (2H, t, J=7.1 Hz), 2.66(2H, t, J=7.1 Hz), 2.51 (2H, t,J=7.1 Hz).

0.9 gr, 6 mM, hydrocinnamic acid and 0.6 gr, 6 equivalents, triphosgenin 30 ml dichloromethane and 1 ml triethyl amine were stirred for 10minutes at room temperature and 0.6 gr ethanol amine were added. After 2hours at room temperature, workup (HCl) gave a viscous oil which wasrecrystalyzed with hexane to give 124 mg white solid, 11%, mp-91.

NMR acetone d₆ 7.30 (5H, m), 3.63 (2H, t, J=5.2 Hz), 3.36 (2H, q, J=5.2Hz), 2.97 (2H, t, J=7.3 Hz), 2.50 (2H, t, J=7.3 Hz).

Prepared similar to AV 28, but with the triphosgen method, from 6 mMindole propionic acid, AV 27. Chromatography gave 0.35 gr viscous oil,13% yield.

NMR CDCl₃ 7.95 (1H(br.s), 7.57 (2H, d, J=8.0 Hz), 7.36-6.90 (9H, m),4.36 (2H, t, J=7.1 Hz), 4.17 (2H, q, J=7.0 Hz), 3.5-2.8 (7H, m).

0.65 gr, 3.9 mM, 4-hydroxy hydro cinnamic acid, 15 ml ethylene glycoland 5 ml thionyl chloride were refluxed 3 hours. Workup gave 0.5 gr 61%,oil.

NMR acetone d₆ 7.02, 6.76 (4H, ABq, J=8.6 Hz), 4.28 (2H, t, J=7.1 Hz),3.63 (2H, t, J=7.1 Hz), 2.85, 2.63 (4H, m).

Prepared as AV 33, 3 mM, from indole propionic acid and tyramine.Chromatography and trituration with ethanol-hexane gave 120 mgpink-white solid, 13%.

NMR acetone d₆ 7.57 (2H, d, J=7.8 Hz)), 7.25-6.97 (8H, m), 3.44 (2H, q,J=7.1 Hz), 3.10 (2H, t, J=7.1 Hz), 2.66 (2H, t, J=7.1 Hz), 2.51 (2H, t,J=7.1 Hz).

To 0.7 gr, 5 mM, hydro cinnamic aldehyde and 0.7 gr, 5 mM, tyramine in20 ml ethanol was added 0.4 gr NaBH₄ and the reaction refluxed 1 hour.Workup gave 0.7 gr viscous oil, 55% yield.

NMR acetone d₆ 7.35 (5H, m), 7.15, 6.85 (4H, ABq, J=8.6 Hz), 2.85 (2H,t, J=6.7 Hz), 2.70 (6H, m), 1.80 (2H, quin., J=7.2 Hz).

Example 2 Synthesis of Polyalkylene Glycol Compounds

Polyalkylene glycol compounds were generally synthesized by preparationof the appropriate alcohol compound (e.g., one of the compoundsdescribed in Example 1, or a hydroxylated derivative thereof) and thenconjugation of the alcohol with a polyalkylene glycol (PAG) polymer(e.g., polyethylene glycol (PEG) or polypropylene glycol (PPG)) of thedesired length.

Compound 1, Phenyl Alaninol

1.2 gr, 32 mM, of LiAlH₄ were added to 2.3 gr, 10 mM, phenyl alanineethyl ester HCl in 50 ml dry ether. After stirring for 2 hours at roomtemperature, water and KOH were added and the reaction product wasextracted with ethyl acetate. After evaporation, 0.8 gr of compound 1, alight yellow oil, was obtained.

Compound 1 crystallized on standing. Mp-70.

NMR CDCl₃ 7.30 (5H, m), 3.64 (1H, dd, J=10.5, 3.8 Hz) 3.40 (1H, dd,J=10.5, 7.2 Hz) 3.12 (1H, m), 2.81 (1H, dd, J=13.2, 5.2 Hz), 2.52 (1H,dd, J=13.2, 8.6 Hz) NMR acetone d₆ 7.30 (5H, m), 3.76 (1H, dt) 3.60 (1H,m) 3.30 (1H, t), 2.85 (2H, m). Helv. Chim. Acta, 31, 1617(1948).Biels.-E3, Vol. 13, p 1757.

Compound 2, Tyrosinol

To 3 gr, 12 mM, L-tyrosine ethyl ester HCl in 50 ml dry ether was added1.2 gr 32 mM LiAlH₄. After stirring 3 hours at room temperature, waterand KOH were added and the reaction was extracted with ethyl acetate.Evaporation gave 1.1 gr of a light yellow oil, 54% yield, which onstanding crystallized. mp-85.

NMR CDCl₃ 7.20 (4H, AB q, J=8.6 Hz), 3.50 (2H, m) 3.20 (1H, m), 2.81(2H, m). NMR tyrosine ethyl ester free base CDCl₃ 7.0, 6.56 (4H, AB q,J=8.8 Hz), 4.20 (2H, q, J=7,0 Hz), 3.70, 3.0, 2.80 (3H, 12 line ABXm),1.28. (3H, t, J=7.0 Hz). JACS, 71, 305(1949). Biels.-E3, Vol. 13, p2263.

Compound 3, Tyrptophanol

To 3 gr, 12.9 mM, L-tryptophan methyl ester HCl in 50 ml dry ether wasadded 1.2 gr, 32 mM LiAlH₄. After stirring 6 hours at room temperaturewater and KOH were added and the reaction extracted with ethyl acetate.Evaporation gave 1.23 gr light yellow oil, 50% yield. On standingcrystallized. Mp-65.

NMR CDCl₃ 7.30 (5H, m), 3.64 (1H, dd, J=10.5, 3.8 Hz) 3.40 (1H, dd,J=10.5, 7.2 Hz) 3.12 (1H, m), 2.81 (1H, dd, J=13.2, 5.2 Hz), 2.52 (1H,dd, J=13.2, 8.6 Hz) J. Het. Chem, 13, 777(1976). Biels.-E5, 22, Vol. 12,p 90.

Compound 4, AV 22

0.66 gr 4-hydroxy hydrocinnamic acid and 4 ml thionyl chloride in 30 mlcyclohexane were refluxed for 2 hours. After evaporation, a white solidwas obtained, to which 0.65 gr oil of Compound 1 (4.3 mM) in 30 mldichloromethane and 0.4 ml triethyl amine were added. After stirring for2 hours at room temperature, water and KOH were added in order toneutralize the pH. The reaction product was extracted withdichloromethane. Evaporation gave 0.8 gr of compound 4, light yellowviscous oil. Part of this product was triturated and recrystallized withethanol to give a white solid. Mp-149.

NMR CDCl₃ 7.30-6.9 (9H, m), 3.50 (2H, m) 3.30 (2H, t, J=7.2 Hz) 2.90(3H, m), 2.60 (2H, t, J=7.2 Hz).

Compound 5, AV 57

0.75 gr, 5 mM, hydrocinnamic acid and 4 ml thionyl chloride in 30 mlcyclohexane were refluxed for 2 hours. Evaporation gave a white solid towhich were added 0.83 gr, 5.5 mM, phenyl alaninol in 30 mldichloromethane and 0.5 ml triethyl amine. After stirring 3 hours atroom temperature, water and KOH were added to neutral pH and thereaction was extracted with dichloromethane. Evaporation gave 0.57 gr ofa yellow viscous oil, 40% yield.

NMR CDCl₃ 7.40-7.10 (10H, m), 3.60 (2H, m) 3.35 (2H, t, J=7.2 Hz) 2.95(3H, m), 2.50 (2H, t, J=7.2 Hz).

Compound 6, AV 58

0.66 gr, 4 mM, 4-hydroxy hydrocinnamic acid and 4 ml thionyl chloride in30 ml cyclohexane were refluxed 3 hours. Evaporation gave a light yellowsolid to which were added 0.72 gr, 4.3 mM, tyrosinol in 30 mldichloromethane and 0.5 ml triethyl amine. After stirring 3 hours atroom temperature, water and KOH were added to neutral pH and thereaction was extracted with dichloromethane. Evaporation gave 0.53 grlight yellow viscous oil, 42% yield.

NMR CDCl₃ 7.30, 7.20 (8H, 2 ABq, J=8.6 Hz), 3.40 (2H, m) 3.30 (2H, t,J=7.2 Hz) 2.90 (3H, m), 2.60 (2H, t, J=7.2 Hz).

Compound 7 AV 59

0.45 gr, 3 mM, hydrocinnamic acid and 3 ml thionyl chloride in 30 mlcyclohexane were refluxed for 2 hours. Evaporation gave a light yellowsolid to which were added 0.66 gr, 3.5 mM, tryptophanol in 30 mldichloromethane and 0.4 ml triethyl amine. After stirring 3 hours atroom temperature, water and KOH were added to neutral pH and thereaction was extracted with dichloromethane. Evaporation gave 0.61 grviscous oil, 63% yield.

NMR CDCl₃ 7.50-7.05 (10H, m), 3.65 (2H, m) 3.32 (2H, t, J=7.3 Hz) 2.92(3H, m), 2.53 (2H, t, J=7.3 Hz).

Compound 8, AV 72

0.45 gr, 3 mM, hydrocinnamic acid and 3 ml thionyl chloride in 30 mlcyclohexane were refluxed for 2 hours. Evaporation gave a light yellowsolid to which were added 0.58 gr, 3.5 mM, tyrosinol in 30 mldichloromethane and 0.4 ml triethyl amine. After stirring for 2.5 hoursat room temperature, water and KOH were added to attain neutral pH andthe reaction was extracted with dichloromethane. Evaporation gave 0.57gr light yellow viscous oil, 63% yield.

NMR CDCl₃ 7.40-7.10 (9H, m),3.60 (2H, m) 3.35 (2H, t, J=7.2 Hz) 2.95(3H, m), 2.50 (2H, t, J=7.2 Hz).

Compound 9, AV 73

0.38 gr, 2 mM, 3-indole propionic acid and 2 ml thionyl chloride in 30ml cyclohexane were refluxed for 2 hours. Evaporation gave light yellowsolid to which were added 0.4 gr, 2.6 mM, phenyl alaninol in 30 mldichloromethane and 0.3 ml triethyl amine. After stirring 2.5 hours atroom temperature, water and KOH were added to neutral pH and thereaction was extracted with dichloromethane. Evaporation gave 0.47 grpink solid, 75% yield.

NMR CDCl₃ 7.58 (1H, d, J=8.0 Hz), 7.40 (1H, d, J=8.0 Hz), 7.30-6.9 (8H,m), 3.50 (2H, m) 3.30 (2H, t, J=7.5 Hz), 2.95 (3H, m), 2.70 (2H, t,J=7.5 Hz).

Compound 10

0.3 gr of Compound 4 (AV 22), 0.8 gr, triphenyl phosphine and 0.55 grethyl diazo carboxylate were added to 1 gr of poly(propylene glycol),(average molecular weight ca 1000), in 60 ml dichloromethane. Stirringfor 2 hours at room temperature, evaporation and chromatography gave0.65 gr of Compound 10, Formula VII, as a viscous oil.

Additional Compounds Synthesized from Phenyl Alaninol

These compounds include those represented by the following formula:

This compound can also be represented as Formula A, where R is apolypropylene glycol polymer and N is the total number of polypropylenemonomers in the polymer:

AV 61:

R=PPG (polypropylene glycol) N=7 MW-706

0.3 gr AV 22 (1 mM), 0.8 gr, 3 mM, triphenyl phosphine and 0.55 gr 3.2mM, ethyl diazo carboxylate were added to 1 gr of poly(propylene glycol)(average mol. weight 424, N=7) in 60 ml dichloromethane. After stirringfor 4 hours at room temperature, evaporation and chromatography gave0.55 gr viscous oil, a 73% yield.

NMR CDCl₃ 7.30-6.9 (9H, m), 4.1-3.0 (m), 2.60 (2H, t, J=7.2 Hz), 1.2-1.1(m)

AV 62

R=PPG N=12 MW-996

Was prepared as above from 0.2 gr AV 22 to give 0.3 gr, 46% yield.

AV 60

R=PPG N=17 MW-1286

Was prepared as above from 0.1 gr AV 22 to give 0.2 gr, 48% yield.

AV 63

R=PPG N=34 MW-2274

Was prepared as above from 0.1 gr AV 22 to give 0.25 gr, 34% yield.

Compounds Synthesized from Compound 5, AV 57

AV86

R=PPG N=7 MW-690

Was prepared as above from 0.22 gr AV 57 to give 0.25 gr, 47% yield.

AV 87

R=PPG N=17 MW-1270

Was prepared as above from 0.2 gr AV 57 to give 0.33 gr, 33% yield.

Compounds Synthesized from Compound 9, AV 73

AV76

R=PPG N=7 MW-729

Was prepared similar to AV 61 above from 0.22 gr AV 73 to give 0.23 gr,47% yield.

AV77

R=PPG N=34 MW-2297

Was prepared as above from 0.2 gr AV 73 to give 0.35 gr, 25% yield.

Compounds Synthesized from Tyrosinol Compounds Synthesized from Compound6, AV 58

AV64

R=PPG N=7 MW-722

Was prepared as above from 0.2 gr AV 58 to give 0.21 gr, 46% yield.

AV65

R=PPG N=17 MW-1302

Was prepared as above from 0.23 gr AV 58 to give 0.28 gr, 29% yield.

Compounds Synthesized from Compound 8, AV 72

AV 74

R=PPG N=7 MW-706

Was prepared similar to AV 61, above, from 0.22 gr AV 72 to give 0.26gr, 50% yield.

AV75

R=PPG N=34 MW-2274

Was prepared as above from 0.2 gr AV 72 to give 0.35 gr, 23% yield.

AV 131

R=PPG N=69 MW-4307

Was prepared as above from AV 72 and poly(propylene glycol (average mol.weight 4,000).

Compounds Synthesized from Tyrptophanol Compounds Synthesized fromCompound 7, AV 59

AV 81

R=PPG N=7 MW-729

Was prepared similar to AV 61, above, from 0.22 gr AV 59 to give 0.26gr, 53% yield.

AV 82

R=PPG N=34 MW-2297

Was prepared as above from 0.2 gr AV 59 to give 0.35 gr, 41% yield.

Example 3 Effect of Compounds on Proliferation Responses of Splenocytes

Spleen cells obtained from native or concanavalin A (Con A)-treated micewere examined in-vitro for their response to T-cell-dependent mitogens.Cells were plated in quadruplicate in 96-well, flat-bottom, microtiterplates (4×10⁵ cells/0.2 ml/well) in RPMI-1640 medium supplemented with5% heat inactivated FCS, 2 mM L-glutamine, 100 U/ml penicillin, 100mg/ml streptomycin and various concentrations of compound 10 (100μg/ml-100 μg/ml). All plates were incubated for 48 h in humidifiedatmosphere of 5% CO₂ in air at 37° C., and then pulsed for 18 h with 1mCi ³[H] thymidine. Cells from each microculture were harvested onfiberglass filters using a multiharvester, and incorporatedradioactivity was measured using standard scintillation techniques.

Incubation with compound 10 has led to a significant inhibition in theproliferation of the splenocytes (Table 1).

TABLE 1 Concentration of Compound 10 No Con A Con A 1 Con A 2 100 μg/ml170 598 1380 1 μg/ml 333 986 2156 100 ng/ml 274 880 2489 1 ng/ml 284 7333609 100 pg/ml 271 1280 3481 0 (control) 837 8775 42551

The effect of AV 75, AV 77, AV 82, AV 86, AV, 87, and a combination ofAV 75 and AV 77 on splenocytes was also tested.

Data from the AV 75 experiment is shown in Table 2, and summarized inFIG. 1. Data from the AV 77 experiment is shown in Table 3, andsummarized in FIG. 2. Data from the AV 75+AV 77 experiment is shown inTable 4, and summarized in FIG. 3. A comparison of the experiments isshown graphically in FIG. 4. In all figures herein, “crude” refers tocrude “Antiviran”, an extract from Bacillus subtilis var. indolasus.See, e.g., PCT publication WO 99/03350.

TABLE 2 No Con A Con A Concentration of Cpm Cpm (% of (% of AV 75control Con A control) control) Control 553 30790 100 100 Crude 1 mg/ml405 499 73.2 1.6 AV 75 1 ug/ml 339 1608 61.3 5.2 AV 75 100 ng/ml 324 72558.5 2.3 AV 75 10 ng/ml 167 529 30.2 1.7 AV 75 1 ng/ml 199 339 35.9 1.1AV 75 100 pg/ml 184 803 33.2 2.6 AV 75 10 pg/ml 126 1994 22.7 6.4

TABLE 3 No Con A Con A Concentration of Cpm Cpm (% of (% of AV 77control Con A control) control) control 553 30790 100 100 crude 1 mg/ml405 499 73.2 1.6 AV 77 1 ug/ml 216 678 39 2.2 100 ng/ml 190 705 34.3 2.210 ng/ml 235 1903 42.5 6.1 1 ng/ml 189 1474 34.1 4.7 100 pg/ml 242 500743.7 16.2 10 pg/ml 169 4427 30.5 14.3

TABLE 4 No Con A Con A Concentration of Cpm Cpm (% of (% of AV 75 + AV77 control Con A control) control) control 553 30790 100 100 crude 1mg/ml 405 499 73.2 1.6 AV 75 + 77 1 ug/ml 182 1439 32.9 4.6 100 ng/ml250 1523 45.2 4.9 10 ng/ml 149 240 26.9 0.7 1 ng/ml 206 2297 37.2 7.4100 pg/ml 181 942 32.7 3 10 pg/ml 101 1420 18.2 4.6

Additional AV 75 experiments were performed. Data from these AV 75experiments is shown in Tables 5 and 6, and is summarized in FIGS. 5 and6.

TABLE 5 No Con A Con A Concentration of Cpm Cpm (% of (% of AV 75control Con A control) control) CONTROL 1433 52111 100 100 CRUDE 1 mg/ml132 176 9.2 0.33 AV 75 old 100 ug/ml 254 189 17.7 0.36 10 ug/ml 302 25021 0.47 1 ug/ml 302 348 21 0.66 0.1 ug/ml 198 220 13.8 0.42 0.01 ug/ml370 273 25.8 0.52 0.001 ug/ml 285 766 19.8 1.47

TABLE 6 No Con A Con A Concentration of Cpm Cpm (% of (% of AV 75control Con A control) control) CONTROL 1277 32124 100 100 CRUDE 1 mg/ml152 261 11.9 0.81 AV 75 new 100 ug/ml 211 8070 16.5 25.1 10 ug/ml 1964054 15.3 12.6 1 ug/ml 350 10300 27.4 32 0.1 ug/ml 349 14365 27.3 44.70.01 ug/ml 293 17536 22.9 54.5 0.001 ug/ml 347 12088 27.1 37.6

Data from the AV 81 experiment is shown in Table 7, and summarized inFIG. 7. Data from the AV 82 experiment is shown in Table 8, andsummarized in FIG. 8. Data from the AV 86 experiment is shown in Table9, and summarized in FIG. 9. Data from the AV 87 experiment is shown inTable 10, and summarized in FIG. 10.

TABLE 7 No Con A Con A Concentration of Cpm Cpm (% of (% of AV 81control Con A control) control) CONTROL 1270 54504 100 100 CRUDE 1 mg/ml333 212 26.2 0.38 AV 81 100 ug/ml 328 3109 25.8 5.7 10 ug/ml 350 34727.5 0.63 1 ug/ml 286 372 22.5 0.68 0.1 ug/ml 278 2440 21.8 4.47 0.01ug/ml 368 8664 28.9 15.9 0.001 ug/ml 224 310 17.6 0.5

TABLE 8 No Con A Con A Concentration of Cpm Cpm (% of (% of AV 82control Con A control) control) CONTROL 1322 36600 100 100 CRUDE 1 mg/ml185 261 13.9 0.71 AV 82 100 ug/ml 435 507 32.9 1.38 10 ug/ml 254 483118.5 13.2 1 ug/ml 232 8896 17.5 24.3 0.1 ug/ml 121 112 9.1 0.3 0.01ug/ml 164 6494 12.4 17.7 0.001 ug/ml 264 11259 19.9 30.7

TABLE 9 No Con A Con A Concentration of Cpm Cpm (% of (% of AV 86control Con A control) control) CONTROL 1576 62506 100 100 CRUDE 1 mg/ml367 296 23.2 0.47 AV 86 100 ug/ml 569 27052 36.1 43.2 10 ug/ml 755 2502547.9 40 1 ug/ml 843 26989 53.4 43.1 0.1 ug/ml 421 15482 26.7 24.7 0.01ug/ml 526 16304 33.3 26 0.001 ug/ml 270 3097 17.1 4.9

TABLE 10 No Con A Con A Concentration of Cpm Cpm (% of (% of AV 87control Con A control) control) CONTROL 1502 50842 100 100 CRUDE 1 mg/ml336 273 22.3 0.53 AV 87 ug/ml 595 39682 39.6 78 10 ug/ml 668 25463 44.450 1 ug/ml 399 12297 26.5 24.1 0.1 ug/ml 142 240 9.4 0.47 0.01 ug/ml 30614795 20.3 29.1 0.001 ug/ml 491 13872 32.6 27.2

In vitro testing with these compounds clearly shows and inhibitoryeffect on splenocyte proliferation at doses ranging between 1 pg/ml and100 ng/ml, and above.

A similar effect was shown with exposure of PHA(phytohemagglutinin)-treated human PMBC cells with AV 74, AV 75, AV 76,a These experiments were performed as the splenocyte experiments. Theresults show a marked inhibitory effect of AV 75 and AV 77, reaching themaximum of inhibition at concentrations of 100 and 10 mg/ml andexhibiting a dose related inhibition at lower concentrations.

Data from the AV 74 experiment is shown in Table 11, and summarized inFIG. 11. Data from the AV 75 experiment is shown in Table 12, andsummarized in FIG. 12. Data from the AV 76 experiment is shown in Table13, and summarized in FIG. 13. Data from the AV 77 experiment is shownin Table 14, and summarized in FIG. 14.

TABLE 11 No PHA PHA Concentration of Cpm Cpm (% of (% of AV 74 controlPHA control) control) CONTROL 701 62309 100 100 CRUDE 1 mg/ml 337 279 480.44 100 ug/ml 363 10243 51.7 16.4 10 ug/ml 270 1972 38.5 3.16 1 ug/ml575 15929 82 25.5 0.1 ug/ml 237 12754 33.8 20.4 0.01 ug/ml 431 5298561.4 85

TABLE 12 No PHA PHA Concentration of Cpm Cpm (% of (% of AV 75 controlPHA control) control) CONTROL 464 49186 100 100 CRUDE 1 mg/ml 272 36058.6 0.73 100 ug/ml 309 781 66.5 1.58 10 ug/ml 394 17516 84.9 35.6 1ug/ml 352 23037 75.8 46.8 0.1 ug/ml 266 11662 57.3 23.7 0.01 ug/ml 27931628 60.1 64.3

TABLE 13 No PHA PHA Concentration of Cpm Cpm (% of (% of AV 76 controlPHA control) control) CONTROL 446 53071 100 100 CRUDE 1 mg/ml 109 28224.4 0.53 100 ug/ml 315 8233 70.6 15.5 10 ug/ml 316 6305 70.8 11.8 1ug/ml 237 6084 53.1 11.4 0.1 ug/ml 238 10634 53.3 20 0.01 ug/ml 20711101 46.4 20.9

TABLE 14 No PHA PHA Concentration of Cpm Cpm (% of (% of AV 77 controlPHA control) control) CONTROL 405 54667 100 100 CRUDE 1 mg/ml 103 35225.4 0.64 100 ug/ml 128 602 31.6 1.1 10 ug/ml 151 1672 37.2 3 1 ug/ml123 4456 30.3 8.15 0.1 ug/ml 209 11424 51.6 20.89 0.01 ug/ml 169 1051841.7 19.24

Example 4 Effect of Compounds on Proliferation of T cells

T cells were isolated from buffy coats (BC) of consenting normal humandonors (Hadassah Hospital Blood Bank). The BC preparations were diluted1:4 with phosphate-buffered saline (PBS) that contained 10 U/mL heparin.Peripheral blood mononuclear cells were separated by Ficoll/Paquedensity centrifugation. Monocytes and B cells were depleted by plasticadherence and passage through nylon wool columns, respectively. Small Tlymphocytes were harvested from the pellet of a discontinuous Percollgradient. The cells were found to be >80% CD3+ by FACS analysis. Cellswere cultured in the presence of various concentrations of compoundand/or phytohemaglutinin (PHA) (1 μg/ml) T cell mitogen. Proliferationwas measured by culturing 1×10⁵ cells in each well of a 96-wellflat-bottomed microtiter plates. 48 hrs and 7 days following addition ofcompound, 1 μCi ³[H] thymidine was added to each well and the cultureswere incubated for an additional 24 hrs. Samples were harvested andincorporated radioactivity was measured. A 7 day incubation withcompounds led to a significant increase in the proliferation ofPHA-stimulated T cells versus PHA alone.

The effect of AV 74, AV 75, AV 76, and AV 77 on PHA(phytohemagglutinin)-treated human peripheral blood mononuclear cells(PMBCs) was investigated. Data from the AV 74 experiment is shown inTable 15, and summarized in FIG. 15. Data from the AV 75 experiment isshown in Table 16, and summarized in FIG. 16. Data from the AV 76experiment is shown in Table 17, and summarized in FIG. 17. Data fromthe AV 77 experiment is shown in Table 18, and summarized in FIG. 18.

TABLE 15 No PHA PHA Concentration of Cpm Cpm (% of (% of AV 74 controlPHA control) control) CONTROL 620 98824 100 100 CRUDE 1 mg/ml 254 37440.9 0.37 100 ug/ml 347 57433 55.9 58.1 10 ug/ml 513 58806 82.7 59.5 1ug/ml 348 49615 56.1 50.2 0.1 ug/ml 365 53942 58.8 54.5 0.01 ug/ml 22433259 36.1 33.65

TABLE 16 No PHA PHA Concentration of Cpm Cpm (% of (% of AV 75 controlPHA control) control) CONTROL 341 99119 100 100 CRUDE 1 mg/ml 156 29545.7 0.29 100 ug/ml 170 229 49.8 0.23 10 ug/ml 309 508 90.6 0.51 1 ug/ml298 27972 87.4 28.22 0.1 ug/ml 209 32316 61.3 32.6 0.01 ug/ml 226 4217862.2 42.55

TABLE 17 No PHA PHA Concentration of Cpm Cpm (% of (% of AV 76 controlPHA control) control) CONTROL 483 105437 100 100 CRUDE 1 mg/ml 189 33939.1 0.32 100 ug/ml 376 67065 77.8 63.6 10 ug/ml 334 58839 69.1 55.8 1ug/ml 463 55382 95.8 52.52 0.1 ug/ml 342 49430 70.8 46.8 0.01 ug/ml 32358054 66.8 55

TABLE 18 No PHA PHA Concentration of Cpm Cpm (% of (% of AV 77 controlPHA control) control) CONTROL 427 95743 100 100 CRUDE 1 mg/ml 126 35129.5 0.36 100 ug/ml 259 283 60.6 0.29 10 ug/ml 137 3593 32 3.75 1 ug/ml234 26456 54.8 27.6 0.1 ug/ml 272 28229 63.7 29.48 0.01 ug/ml 284 3142266.5 32.8

Example 5 Immunosuppressive Effect of Compounds

Human T cells were isolated as described above. The immunosuppressiveeffect of AV 72, AV 72, AV 74, AV 75, AV 76, AND AV 77 on PMBCs (bothnon-activated and ConA-activated) relative to cells treated with CsA(cyclosporin A) is shown in FIGS. 19, 20, 21, 22, and 23, respectively.The effect of AV 61, relative to AV 75 and AV 77 is shown in FIG. 24.The results dearly show the immunosuppressive effect of AV 75 and AV 77on PBMCs, reaching the maximal suppressive effect with 100 ng/ml with AV75 and 100 pg/ml with AV 77.

The immunosuppressive effect of AV 61, AV 75, and AV 77 on dendriticcells (both non-activated and ConA-activated) relative to cells treatedwith CsA (cyclosporin A) is shown in FIG. 25. Thus, the suppressiveeffect of AV 77 was shown not only on the PBMCs but also on antigenpresenting cells (APCs) such as dentritic cells, reaching a suppressiveeffect of 65% with 1 pg/ml).

Example 6 Effect of AV 75 on Experimental AutoImmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) is an organ-specific Tcell-mediated autoimmune disease resulting in demyelination of the whitematter in the central nervous system (CNS). In many of its clinical andhistopathological aspects, EAE resembles human multiple sclerosis (MS)and acute disseminating encephalomyelitis. Early histopathologicalmanifestations of the disease are infiltrating monocytic lesionsfollowed by infiltrating lymphocytic ones in the brain and spinal cord,with areas of demyelination in the white matter of the CNS. The abilityof AV 75 to abrogate pathogenesis of acute EAE was investigated.

Animals:

Twenty-four female SJL/J mice, 6-12 week old were used for thisexperiment. They were housed under standard conditions in top filteredcages, and were fed a regular diet without antibiotics.

Induction and Clinical Evaluation of EAE:

Induction of acute EAE in SJL mice was based on a modification ofBernard's procedure (Bernard et al., J. Immunol. 114, 1537-1540 (1975)).Briefly, equal volumes of Mouse Spinal Cord Homogenate (MSCH) 100 mg/mlin PBS and CFA enriched with Mycobacterium tuberculosis H37Ra (6 mg/ml)were emulsified. The emulsion (50-100 μl) was administeredsubcutaneously into the four footpads of each mouse. Immediatelythereafter, and 2 days later, mice were injected i.v. with pertussigen.All animals were examined daily for signs of disease. The first clinicalindication appeared on day 11-12 post immunization and were scoredaccording to the following six point scale: 0, no abnormality; 1, mildtail weakness; 2, tail paralysis; 3, tail paralysis and hind legparesis; 4, hind leg paralysis or mild forelimb weakness; 5,quadriplegia or moribund state; 6, death.

AV 75 Treatment:

Av 75 was administered intraperitoneally at two dose levels of 1 μg and100 μg per animal, 3 times a week for the duration of the experiment.The administration was initiated on day 0, immediately following theimmunization with MSCH for the induction of EAE. AV 75 administered as100 pg/ml in drinking water of mice (ad libitum), changing the watertwice a week has also resulted in positive results in the acute EAEmodel of MS.

In vitro Proliferative Responses of Lymphocytes:

Draining lymph nodes were excised from EAE and control mice at day 10after immunization. Single cell suspensions of lymphocytes were assayedby [³H]thymidine incorporation. The assay was carried out by seeding4×10⁵ cells/well in 0.2 ml of RPMI medium supplemented with 2.5% fetalcalf serum, L-glutamine, antibiotics and optimal concentrations of thefollowing antigens: PLP (proteolipid protein, a myelin peptide), PPD(tuberculin-purified protein derivative), ConA (concanavalin A), SEB(Staphylococcal enterotoxin B) MOG (myelin oligodendrocytesglycoprotein) and LPS (lipopolysaccharide).

Results:

The 24 mice were divided in 3 groups: 8 mice served as control; 8 micereceived 1 μg per animal and 8 mice received 100 μg per animal on eachinjection, three times a week during the experiment. On day 10, two micefrom each group were sacrificed and their draining lymph nodes wereexcised with the aim to be assayed in the in vitro proliferation assayfor response to antigens.

The remaining 6 animals per group continued with the experiment. Oneanimal in the 100 μg group died for unknown reasons. The results of theexperiment are presented in FIGS. 26A, B, and C: the control groupdeveloped the disease as expected. Only 2 animals in each AV 75 treatedgroup developed relatively mild signs of the disease, and in the graphthe differences between the treated and untreated groups can be clearlyseen.

The results of the proliferative response to antigens of the lymphocytesobtained from the draining lymph nodes are summarized in FIG. 27A. Datafrom individual antigen experiments is presented in FIGS. 27B-F. Datafrom the control (no antigen) experiment is shown in FIG. 27G. Theseresults represent the average of the 2 lymph nodes from each group. Theinhibitory effect of the 2 doses of AV 75 against all the antigenstested is evident. Especially remarkable is the effect against the PLPantigen, the most distinctive antigen in EAE.

Example 7 Effect of AV 75 and AV 77 on Transplant Rejection

The effect of AV 75 and AV 77 transplant rejection was investigated invivo in a rat acute renal allograft rejection model. Kidneys fromBrown-Norway rats (donors) were orthotopically transplanted into Lewisrats (recipients), followed by right nephrectomy of the recipients. Inthe group receiving two injections of 100 ng per rat of AV 75, on days 0and 4 after transplantation, prevention of the rejection has beenobserved for long periods (e.g., up to 53 days as compared to up to 4days survival of the controls). Data is summarized in Table 19.

TABLE 19 Survival Treatment Subject Compound time transplantation andTAA1 AV75 2 days compound (100 ug) TAA2 AV75 4 days TAA3 AV75 4 daysTAB1 AV77 3 days TAB2 AV77 3 days TAB3 AV77 2 days TAC1 vehicle 3 daysTAC2 vehicle 2 days TAD1 AV75 3 days TAD2 AV75 3 days TAD3 AV75 2 daysTAE1 AV77 3 days TAE2 AV77 3 days TAE3 AV77 2 days TAF1 vehicle 3 daysTAF2 vehicle 3 days transplantation and TAA1 AV75 4 days compound (100ng) TAA2 AV75 6 days TAA3 AV75 5 days TAB1 AV77 4 days TAB2 AV77 4 daysTAB3 AV77 2 days TAB4 AV77 4 days TAC1 vehicle 3 days TAC2 vehicle 4days TAC3 vehicle 3 days TAC4 vehicle 3 days TAD1 AV75 53 days TAD2 AV755 days TAD3 AV75 35 days TAD4 AV75 34 days TAE1 AV77 3 days TAE2 AV77 4days TAE3 AV77 3 days TAE4 AV77 5 days TAF1 vehicle 4 days TAF2 vehicle4 days TAF3 vehicle 3 days TAF4 vehicle 3 days transplantation and TAA1AV75 4 days compound (100 pg) TAA2 AV75 6 days TAA3 AV75 7 days TAA4AV75 6 days TAB1 AV77 3 days TAB2 AV77 4 days TAB3 AV77 6 days TAC1vehicle 3 days TAC2 vehicle 3 days TAC3 vehicle 3 days TAD1 AV75 3 daysTAD2 AV75 10 days TAD3 AV75 8 days TAE1 AV77 5 days TAE2 AV77 4 daysTAE3 AV77 5 days TAF1 vehicle 3 days TAF2 vehicle 4 days TAF3 vehicle 3days transplantation and TAN1 AV75 6 days compound (100 pg) TAN2 AV75 5days i.p. day 0 and day 3 TAN3 AV75 6 days TAN4 AV75 7 days TAN5 AV75 6days TAN6 AV75 5 days transplantation and TAG1 AV75 3 days compound (100ng) TAG2 AV75 4 days i.p. day 0 and day 3 TAG3 AV75 3 days TAG4 AV75 3days TAG5 AV75 4 days transplantation and TAH1 AV75 4 days compound (100ng) TAH2 AV75 3 days i.p. day −1 and day 3 TAH3 AV75 4 days TAH4 AV75 3days transplantation and TAI1 AV75 4 days compound (100 ng) i.p. day 0and day 4 transplantation and TAJ1 AV75 3 days compound (100 ng) i.p.day −1 and day 3

It will be appreciated that the present invention is not limited by whathas been described hereinabove and that numerous modifications, all ofwhich fall within the scope of the present invention, exist. Rather thescope of the invention is defined by the claims that follow.

1. A compound of formula I:

wherein: the dotted line represents a single or a double bond; R₁ and R₂are the same or different and, independently of each other, are groupsrepresented by the formula:

in which R₅ is H, OH or OR₆, where R₆ is a linear or branched C₁-C₄alkyl; A-B is a group represented by the formula:

 and X is, —CH₂O—, —CH₂CH₂O—, —CH(CH₃)CH₂O— or —CH₂CH(CH₃)O—; Z is—CH₂CH₂O—, —CH(CH₃)CH₂O— or —CH₂CH(CH₃)O—; m is 1, and n is an integerof 1-50 and salts thereof.
 2. The compound of claim 1, wherein R₁ is:

where R₅ is H or OH.
 3. The compound of claim 2, wherein R₁ is phenyl.4. The compound of claim 2, wherein R₁ is:


5. The compound of claim 1, wherein R₂ is:

wherein R₅ is H or OH.
 6. The compound of claim 5, wherein R₂ is phenyl.7. The compound of claim 5, wherein R₂ is


8. The compound of claim 1, wherein X is —CH₂O—.
 9. The compound ofclaim 1, wherein Z is —CH(CH₃)CH₂O— or —CH₂CH(CH₃)O—.
 10. The compoundof claim 8, wherein Z is —CH(CH₃)CH₂O— or —CH₂CH(CH₃)O—.
 11. Thecompound of claim 8, wherein R₁ is


12. The compound of claim 10, wherein R₁ is


13. The compound of claim 8, wherein R₁ is


14. The compound of claim 10, wherein R₁ is


15. The compound of claim 8, wherein R₂ is


16. The compound of claim 10, wherein R₂ is


17. The compound of claim 8, wherein R₂ is


18. The compound of claim 10, wherein R₂ is


19. The compound of claim 1, wherein n is an integer of 1-20.
 20. Thecompound of claim 8, wherein n is an integer of 1-20.
 21. The compoundof claim 10, wherein n is an integer of 1-20.
 22. The compound of claim1, wherein n is an integer of 10-20.
 23. The compound of claim 8,wherein n is an integer of 10-20.
 24. The compound of claim 10, whereinn is an integer of 10-20.
 25. The compound of claim 1, wherein n is 7.26. The compound of claim 8, wherein n is
 7. 27. The compound of claim10, wherein n is
 7. 28. The compound of claim 1, wherein n is an integerof 5-50.
 29. The compound of claim 8, wherein n is an integer of 5-50.30. The compound of claim 10, wherein n is an integer of 5-50.
 31. Apharmaceutical composition comprising one or more compounds according toclaim 1 together with one or more pharmaceutically acceptable excipientsor adjuvants.
 32. The composition of claim 31, formulated as uncoatedtablets, coated tablets, pills, capsules, powder or suspension.
 33. Thecomposition of claim 31, formulated in an ointment, cream or gel form.34. A compound of formula II:

wherein R is a polyalkylene glycol polymer having n units, where n is aninteger from 1-50, and R₇ and R₈ are selected from the combinationsshown: R₇ R₈

and salts thereof.
 35. A compound of the formula:

wherein Z is —CH(CH₃)CH₂O— or —CH₂CH(CH₃)O—, or salt thereof.
 36. Apharmaceutical composition comprising a compound of the formula:

wherein, Z is —CH(CH₃)CH₂O— or —CH₂CH(CH₃)O—, together with one or morepharmaceutically acceptable excipients or adjuvants.